Improved mycobacterial protein production using a Mycobacterium smegmatis groEL1ΔC expression strain.

ABSTRACT:

Background

The non-pathogenic bacterium Mycobacterium smegmatis is widely used as a near-native expression host for the purification of Mycobacterium tuberculosis proteins. Unfortunately, the Hsp60 chaperone GroEL1, which is relatively highly expressed, is often co-purified with polyhistidine-tagged recombinant proteins as a major contaminant when using this expression system. This is likely due to a histidine-rich C-terminus in GroEL1.

Results

In order to improve purification efficiency and yield of polyhistidine-tagged mycobacterial target proteins, we created a mutant version of GroEL1 by removing the coding sequence for the histidine-rich C-terminus, termed GroEL1ΔC. GroEL1ΔC, which is a functional protein, is no longer able to bind nickel affinity beads. Using a selecti

SUBMITTER: Noens EE 

PROVIDER: S-EPMC3076238 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature